N-acyl-phosphatidyl-ethanolamines and/or mixtures of n-acyl-ethanolamines with phosphatidic acids or lysophosphatidic acids

ABSTRACT

Pharmaceutical, cosmetic and dietetic compositions and functional foods, constituted by: A) phospholipid mixtures containing N-acyl-phosphatidyl-ethanolamines (NAPEs) and/or B) phospholipid mixtures containing N-acyl-ethanol amines (NAEs) together with phosphatidic acids (PAs) and/or lysophosphatidic acids (LPAs) with the proviso that said N-acyl-phosphatidyl-ethanolamines (NAPEs) do not include N-oleoyl-phosphatidyl-ethanolamine. as well as new phosphobioflavonic complexes of NAPE or NAE with one or more bioflavonoids, are disclosed.

The present invention relates to pharmaceutical, cosmetic and dieteticcompositions and functional foods, constituted by:

-   -   A) phospholipid mixtures containing        N-acyl-phosphatidyl-ethanolamines (NAPEs);

and/or

-   -   B) phospholipid mixtures containing N-acyl-ethanolamines (NAEs)        together with phosphatidic acids (PAs) and/or lysophosphatidic        acids (LPAs), with the proviso that said        N-acyl-phosphatidyl-ethanolamines (NAPEs) do not include        N-oleoyl-phosphatidyl-ethanolamine.

Also disclosed are new phosphobioflavonic complexes of NAPE or NAE plusPA and/or LPA, with one or more bioflavonoids.

N-Acyl-ethanolamines (NAEs) and N-acyl-phosphatidyl-ethanolamines(NAPEs) are known to be present in many foods of animal and vegetableorigin (H. H. Schmid et al., 1990, Prog. Lipid Res., 29, 1-43), and areparticularly abundant in foods such as soy, eggs and chocolate (K. D.Chapman et al., 1993, Arch. Biochem. Biophys, 301, 21-23; E. Di Tomasoet al., 1996, Nature, 382, 677-678).

The NAEs are formed in vivo by hydrolysis of a NAPE molecule that givesrise to a mixture of NAE and a molecule of phosphatidic acid (PA) which,in turn, can be hydrolysed to lysophosphatidic acid (LPA) in accordancewith the following scheme 1.

GB 2051069 discloses the anti-lipemic and anti-atherosclerotic activityof N-Oleoyl-phosphatidylethanolamine (NOPE) and excludes any significantactivity of other N-acyl-derivatives.

NAEs have also been known for some time for their interestingpharmacological properties: N-arachidonoyl-ethanolamine has beendemonstrated in vitro to be a cannabinoid receptor agonist (L. Hanus,1993, J. Med. Chem., 36, 3032-3034); N-palmitoyl-ethanolamine, whenadministered intraperitoneally to rats, possesses anti-inflammatory andanti-anaphylactic activity (L. Facci et al., 1995, Proc. Natl. Acad.Sci. USA, 92, 3376-3380); N-palmitoyl-ethanolamine andN-stearoyl-ethanolamine have proved useful in the pharmacologicaltreatment of inflammatory disorders resulting from degranulation of themast cells (EP-A-0550006); they also inhibit peroxidation of themitochondrial membranes in vitro (N. M. Gulaya et al., 1998, Chem. Phys.Lipids, 97, 49-54); N-oleoyl-ethanolamine (NOE) has a significantanorexic effect in the rat, when administered by the intraperitonealroute (F. Rodriguez de Fonseca et al., 2001, Nature, 414, 209-212).Since it is well known that NAEs are easily hydrolised to free fattyacids and ethanolamine in the gastrointestinal tract, its activity bythe oral route is not expected.

The present invention relates to pharmaceutical and dieteticcompositions and functional foods, constituted by:

-   -   A) phospholipid mixtures containing        N-acyl-phosphatidyl-ethanolamines (NAPEs);

and/or

-   -   B) phospholipid mixtures containing N-acyl-ethanolamines (NAEs)        together with phosphatidic acids (PAs) and/or lysophosphatidic        acids (LPAs),

with the proviso that said N-acyl-phosphatidyl-ethanolamines (NAPEs) donot include N-oleoyl-phosphathidyl-ethanolamine.

The structural formulas of NAE, PA and LPA are shown in scheme 2,wherein R₁, R₂ and R₄ are acyl residues of long-chain fatty acids, inparticular residues of palmitic, stearic, oleic, linoleic, conjugatedlinoleic, linolenic, gamma-linolenic, eicosapentaenoic anddocosahexanoic acids, etc.

The phospholipid mixtures may be present in the compositions of theinvention in the form of their complexes with bioflavonoids. Saidcomplexes, hereinafter called “phosphobioflavonic complexes”, are afurther object of the invention.

Complexes of phospholipids ouch as lecithins, phosphatidylcholine,phosphatidylethanolamine and phosphatidylserine with a number of plantextracts have been disclosed (U.S. Pat. No. 4,963,527, U.S. Pat. No.4,895,839, EP 283713). Said complexes are reported to increase thebioavailability of the plant extract. In the phosphobioflavoniccomplexes of the invention, NAPE or NAE plus PA and/or LPA provide anunexpected synergism for the considered applications and do not actmerely as carriers of bioflavones.

Said complexes, constituted by aggregation of the phospholipid activecomponents (NAPE and/or NAE plus PA and/or NAE plus LPA) with one ormore types of bioflavonoids, can be obtained by suspending a dryphospholipid residue under strong stirring for a few minutes at atemperature preferably between 40° C. and 65° C. in a hydroalcoholicsolution (alcohol preferably between 70 and 90%), buffered to an acid pH(pH preferably between 3 and 5), containing a fraction of one or moretypes of bioflavonoids, preferably in a percentage of between 0.5 and15% by weight of the hydroalcoholic solution. When stirring isinterrupted, ethanol is partially evaporated from the resulting emulsionunder vacuum and then dehydrated by spray drying, to produce a drygranular residue of phosphobioflavonic complexes.

Examples of bioflavonoids which can be used to produce thesephosphobioflavonic complexes include:

-   -   a) simple polyphenols such as cinnamic, cumaric, caffeic and        ferulic acids;    -   b) flavones such as hesperidin, naringenin and taxifolin;    -   c) flavonols such as kaempferol glycoside, quercetin, quercetin        glycoside, myricetin and myricetin glycoside;    -   d) isoflavones such as genistein and daidzein;    -   e) proanthocyanidins such as procyanidin B1, procyanidin B2,        procyanidin B3 and procyanidin C-1;    -   f) anthocyanidins such as pelargonidin, delphinidin, malvidin        and petunidin;    -   g) catechins such as epicatechin, epicatechin gallate,        epigallocatechin, catechins and gallocatechins;    -   h) tannins.

As mentioned, molecules of NAPE, NAE, PA and LPA are naturally presentin the lipid fractions of many foodstuffs normally used in the humandiet (soy lecithins, eggs, cocoa, meat, oily extracts of various seeds,etc.), and can easily be extracted and isolated to various degrees ofpurity in accordance with conventional methods. Alternatively, the NAPEand NAE molecules can be obtained by synthesis according to chemicalprocesses which have been known for some time.

NAE can prepared from ethanolamine and the corresponding fatty acid, forexample in accordance with the methods described in:

-   -   Roc E. T. et al. (1952), J. Am. Chem. Soc., 74, 3442    -   Chandrakumar N. S. et al. (1982), Biochim. Biophys Acta, 711,        357.

NAPE can prepared from phosphatidylethanolamine and the correspondingfatty acid chloride or anhydride, in accordance with the methodsdescribed in:

-   -   Schmid P. C. et al. (1988), J. Biol. Chem., 288 (6), 9802    -   Epps D. E. et al. (1980), Biochim. Biophys Acta, 618, 420    -   GB 2051069 B.

Another method for the preparation of NAPE by means of the enzymephospholipase D, disclosed in U.S. Pat. No. 4,783,402, is illustrated inthe scheme below:

wherein

R₁, R₂ and R₄ represent the alkyl chain of saturated, mono- orpolyunsaturated fatty acids with 12-22 C atoms;

R₃ represents a residue of choline, ethanolamine, inositol, glycerol,serine.

The therapeutically effective doses of preparations based on NAPE and/orNAE plus PA and/or LPA vary:

-   -   a) in the case of NAPE from 0.5 to 50 mg, and preferably from 1        to 10 mg/day per kg of body weight;    -   b) in the case of NAE plus PA and/or LPA from 0.5 to 100 mg, and        preferably from 2 to 20 mg/day/kg of body weight. In this        mixture of NAE+PA and/or LPA, the percentage of NAE can vary        between 1 and 70%, and preferably between 25 and 50% of the        total co-mixed lipids.

The compositions of the invention may also contain other nutritionalcomponents which further implement the therapeutic properties andbenefits of NAPE and/or the mixtures of NAE+PA and/or LPA. Examples ofthese components are:

-   -   a) vitamins and vitamin-like factors such as vitamin E, vitamin        C, β-carotenes, vitamin A, vitamin D, lipoic acid and CoQ;    -   b) extracts of vegetables and/or medicinal plants based on mono-        and diterpenes, saponins, and phytosterols;    -   c) proteins, peptides or aminoacids and their derivatives such        as glutathione, carnosine, arginine, glutamine, carnitine,        creatine and taurine;    -   d) trace elements and mineral salts such as Ca, Mg, Cr, Se, Va,        Zn and Cu;    -   e) mixtures of natural amphiphilic detergents such as        phospholipids and lysophospholipids; glycolipids; amphiphilic        proteins; mono- and diglycerides; bile acids or salts able to        incorporate NAPE and/or mixtures of NAE+PA and/or LPA in lipid        emulsions of various types which help to increase their        absorption and bioavailability in vivo.

The active components, stored as dehydrated granulates or powders, canbe used as such or in the form of aqueous or oily solutions to makevarious galenical preparations such as gelatin capsules, tablets,dragees, sachets, effervescent and non-effervescent cachets, chewinggum, etc.

Said active components in the form of dehydrated granulates or powderscan also be used to make various functional foods:

-   -   a) mixed with oils to make sundry dressings, sauces, creams,        mayonnaise, etc;    -   b) mixed with flour to make bread, pasta, crackers, biscuits and        other baked products;    -   c) added to fruit juices and squashes, mineral waters, soft        drinks and other drinks;    -   d) added to milk and derivatives thereof (yoghurt, flans,        ricotta and cheese).

The pharmaceutical or dietetic compositions of the invention have provedsurprisingly active in:

-   -   a) controlling excess weight and consequently reducing the risks        connected with excess weight and obesity;    -   b) improving the functionality of the mitochondria and the        production of cell energy;    -   c) increasing the antioxidant defences in the various tissues;    -   d) improving the “fluidity” of the cell membranes and        consequently the functionality of the membrane proteins        (enzymes, receptors, carriers of essential nutrients and trace        elements, etc.).

The preparations of the invention can therefore be used as adjuvants inthe treatment of aging and many metabolic disorders connected with it(obesity and excess weight; diabetes; cerebro-degenerative disorderssuch as Alzheimer's disease, Parkinson's disease and senile dementia;stress, depression; tumours; menopausal syndromes; osteoporosis;prostate hypertrophy; skin aging; panniculopathy (cellulitis); andalopecia), possibly in combination with known drugs or diet supplements.

The invention therefore also concerns the use of phospholipid mixturescontaining

-   -   A) N-acyl-phosphatidyl-ethanolamines (NAPEs);

and/or

-   -   B) phospholipid mixtures containing N-acyl-ethanolamines (NAEs)        together with phosphatidic acids (PAs) and/or lysophosphatidic        acids (LPAs)        for the preparation of medicaments having anorexic activity or        of medicaments or foodstuffs for the treatment of aging, obesity        and excess weight; diabetes; cerebro-degenerative disorders such        as Alzheimer's disease, Parkinson's disease and senile dementia;        stress, depression; tumours; menopausal syndromes; osteoporosis;        prostate hypertrophy; skin aging; panniculopathy (cellulitis)        and alopecia.

The invention is illustrated in greater detail in the followingexamples.

EXAMPLE 1

98 g of N-linoleoyl-phosphatidylethanolamine+1 g of d-α-tocopherol+1 gof lipoic acid.

The various compounds are dissolved and mixed in 10 volumes ofchloroform: methanol (2:1, vol/vol). The solvent is evaporated undervacuum, and the resulting dry residue is re-suspended in an aqueoussolution buffered to physiological pH to form an aqueous mixture of aphospholipid emulsion containing the active component(N-linoleoyl-phosphatidylethanolamine). The aqueous mixture can befrozen and dehydrated to obtain a dry residue of the phospholipid activecomponent.

EXAMPLE 2

20 g of N-eicosapentaenoyl-ethanolamine+60 g of phosphatidic acid+80 gof soy phosphatidylcholine+1 g of d-α-tocopherol+1 g of lipoic acid.

The various compounds are dissolved in chloroform-methanol and treatedas described in example 1 to obtain an aqueous mixture of a phospholipidemulsion containing the active components(N-eicosapentaenoyl-ethanolamine and phosphatidic acid). The aqueousmixture can be frozen and dehydrated to obtain a dry phospholipidresidue of the active components as described in example 1.

EXAMPLE 3

20 g of N-linolenoyl-ethanolamine+40 g of lysophosphatidic acid+1 g ofd-α-tocopherol+1 g of lipoic acid.

The various compounds are dissolved in chloroform-methanol and treatedas described in example 1 to obtain an aqueous mixture of a phospholipidemulsion containing the active components (N-linolenoyl-ethanolamine andphosphatidic acid). The aqueous mixture can be frozen and dehydrated toobtain a dry phospholipid residue of the active components as describedin example 1.

EXAMPLE 4

20 g of N-gamma-linolenoyl-phosphatidylethanolamine+80 g of a mixture oflysophospholipids (45% lysophosphatidylcholine+35%lysophosphatidylethanolaniine+20% lysophosphatidylinositol)+1 g ofd-α-tocopherol+1 g of lipoic acid.

The various compounds are dissolved in chlotoform-methanol and treatedas described in example 1 to obtain an aqueous mixture of a phospholipidemulsion containing the active constituent(N-gamma-linolenoyl-phosphatidylethanolamine). The aqueous mixture canbe frozen and dehydrated to obtain a dry phospholipid residue of theactive constituent as described in example 1.

EXAMPLE 5

20 g of a dry phospholipid residue obtained as described in examples 1-4above+200 g of an oily solution (olive oil, soy, corn, sunflower,borage, blackcurrant, fish or seaweed oils, or mixtures thereof).

20 g of dry phospholipid residues is slowly dissolved in 200 g of oilysolution under slow, continuous stirring. The phospholipids of the dryresidues are restructured in the oily solutions to form an oil-dispersedmicellar organisation containing the active components.

EXAMPLE 6

100 g of a dry phospholipid residue ofN-docosahexanoyl-phosphatidylethanolamine, obtained as described inexample 1, is re-suspended under strong stirring for 5 minutes at 45° C.in 900 ml of a hydroalcoholic solution (75% alcohol), buffered to pH4.5, containing 5% by weight of green tea catechins and epicatechins.The resulting emulsion is then cooled to room temperature and dehydratedby spray drying to form a dry granular residue of phosphobioflavoniccomplexes of N-docosahexanoyl-phosphatidylethanolamine and green teacatechins.

EXAMPLE 7

50 g of N-linolenoyl-ethanolamine and 50 g of lysophosphatidic acid(CLPA) are slowly added under strong stirring at 60° C. and emulsifiedfor 10 minutes in 900 ml of a hydroalcoholic solution (85% alcohol)buffered to pH 4.0, containing 10% by weight of a mixture of catechins,epicatechins and proanthocyanidins extracted from grape seeds. Whenstirring is arrested, the resulting emulsion is cooled to roomtemperature and dehydrated by spray drying to form a dry granularresidue of phosphobioflavonic complexes of N-linolenoyl-ethanolamine andgrape-seed bioflavonoids.

Pharmacological and/or Dietetic Tests

A series of experimental tests on rats and clinical tests on man havebeen carried out to study the pharmacological and/or dieteticcharacteristics of the composition of the invention.

In the experimental tests, the rats were given a high-calorie,high-triglyceride, high-cholesterol diet. The following parameters wereevaluated after twenty days treatment:

-   -   1) effect of the compositions on the lipoperoxide levels in the        plasma, liver, brain and heart;    -   2) effect of the compositions on variations in body weight;    -   3) effect of the compositions on variations in membrane fluidity        of ghost erythrocytes and plasma platelets;    -   4) effect of the compositions on the functionality of the        hepatic mitochondria, evaluated by measuring: a) O₂        consumption; b) reduced glutathione; c) the potential of the        mitochondrial membranes;    -   5) effect of the compositions on plasma levels of total        cholesterol and HDL-cholesterol;    -   6) effect of the compositions on plasma levels of total        triglycerides.

80 Male rats weighing 150-200 g each were used. The animals were dividedinto 8 groups of 10 animals:

-   -   1^(st) group: control (C); 10 animals (control at time 0) were        used as is, and 10 were given a standard high-calorie, high-fat,        high-cholesterol diet for 20 days, consisting of:    -   casein: 20%; mixture of trace elements and mineral salts: 3.5%;        mixture of vitamins: 0.1%; choline bitartrate: 0.2%; cellulose:        2%; cholesterol: 0.5%; sodium cholate: 0.25%; saccharose:        58.44%, lard: 10.0% and olive oil: 4.9%.    -   2^(nd) group: treated with N-oleoyl-ethanolamine as such (NOE);        the animals were given the same diet as the controls for 20        days, except that 50 mg of NOE replaced the same quantity of        olive oil (olive oil used: 4.85%).    -   3^(rd) group: treated with N-oleoyl-phosphatidylethanolamine        prepared as described in example 1 (NOPE); the animals were        given the same diet as the controls for 20 days, except that 50        mg of NOPE (prepared as described in example 1) replaced the        same quantity of olive oil (olive oil used: 4.85%).    -   4^(th) group: treated with N-oleoyl-ethanolamine+phosphatidic        acid prepared as described in example 2: (NOE+PA); the animals        were given the same diet as the controls for 20 days, except        that 400 mg of the preparation described in example 2        (containing ˜50 mg of NOE and 150 mg of PA) replaced the same        quantity of olive oil (olive oil used: 4.50%).    -   5^(th) group: treated with        N-oleoyl-ethanolamine+lysophosphatidic acid prepared as        described in example 3: (NOE+LPA); the animals were given the        same diet as the controls for 20 days, except that 150 mg of the        preparation described in example 3 (containing ˜50 mg of NOE and        100 mg of LPA) replaced the same quantity of olive oil (olive        oil used: 4.75%).    -   6^(th) group: treated with “phosphobioflavonic complexes” of        N-oleoyl-phosphatidylethanolamine and green tea bioflavones        (B.F.) prepared as described in example 6 (NOPE+B.F.). The        animals were given the same diet as the controls for 20 days,        except that 50 mg of NOPE and 25 mg of B.F. (corresponding to        ˜75 mg of the preparation described in example 6) replaced the        same quantity of olive oil (olive oil used: 4.825%).

7^(th) group: treated with green tea bioflavones (B.F.). The animalswere given the same diet as the controls for 20 days, except that 25 mgof B.F. replaced the same quantity of olive oil (olive oil used:4.875%). TABLE I Percentage variations in membrane fluidity of ghosterythrocytes and plasma platelets (expressed as a % of the controlvalues at time 0) of the rats before and after 20 day diet treatment.Membrane fluidity Membrane fluidity (ghost erythrocytes) (plasmaplatelets) 1A) Control rats at time 0 100% 100% 1B) Control rats after72% 69% a 20-day diet 2) Treated rats (NOE) 72% 70% 3) Treated rats(NOPE) 84% 81% 4) Treated rats (NOE + 86% 81% PA) 5) Treated rats (NOE +83% 80% LPA) 6) Treated rats (NOPE + 91% 92% B.F.) 7) Treated rats(B.F.) 73% 70%

TABLE II Lipoperoxide levels [expressed as nmoles of malonyldialdehyde(MDA) per gram of tissue or per ml of plasma] in the plasma, livers,brains and hearts of the rats before and after 20 day diet treatment.MDA MDA MDA MDA PLASMA LIVER BRAIN HEART 1A) Control rats 2.5 ± 0.5 25.5± 5.9 55 ± 4 24 ± 5 at time 0 1B) Control rats 5.1 ± 0.6 44.2 ± 8.2 108± 6 45 ± 6 after a 20-day diet 2) Treated rats (NOE) 5.0 ± 0.6 44.1 ±8.2 106 ± 7 44 ± 9 3) Treated rats (NOPE) 3.8 ± 0.4 33.1 ± 6.5  85 ± 932 ± 7 4) Treated rats (NOE + 3.7 ± 0.4 31.8 ± 8.2  88 ± 7 34 ± 9 PA) 5)Treated rats (NOE + 3.0 ± 0.3 33.5 ± 7.8  77 ± 5 34 ± 7 LPA) 6) Treatedrats (NOPE + 2.8 ± 0.3 29.7 ± 6.8  75 ± 4 30 ± 6 B.F.) 7) Treated rats(B.F.) 5.0 ± 0.5 44.0 ± 7.1 105 ± 7 44 ± 7

TABLE III Variation in body weight and total cholesterol, HDLcholesterol and triglyceride levels in the plasma of the rats before andafter 20 day diet treatment. Total HDL Total Body cholesterolcholesterol triglycerides weight (mg dl⁻¹) (mg dl⁻¹) (mg dl⁻¹) (gm) 1A)Control rats 35.6 ± 1.8 26.2 ± 1.4 50.2 ± 7.7 180 ± 12 at time 0 1B)Control rats 126.2 ± 13.5 29.4 ± 1.6 82.5 ± 9.5 224 ± 19 after a 20-daydiet 2) Treated rats 120.4 ± 12.7 28.9 ± 2.8 80.5 ± 6.8 221 ± 16 (NOE)3) Treated rats 110.3 ± 10.1 31.6 ± 3.9 71.4 ± 8.7 209 ± 18 (NOPE) 4)Treated rats 103.9 ± 12.4 29.9 ± 2.0  70.4 ± 10.5 208 ± 14 (NOE + PA) 5)Treated rats 101.7 ± 8.9  32.1 ± 3.8 68.5 ± 7.9 206 ± 20 (NOE + LPA) 6)Treated rats   80 ± 7.5 31.4 ± 3.9 60.2 ± 6.4 191 ± 14 (NOPE + B.F.) 7)Treated rats 123.5 ± 12.4 29.3 ± 1.5 80.7 ± 7.1 218 ± 17 (B.F.)

TABLE IV Variations in hepatocellular oxygen consumption, membranepotential of mitochondria and reduced hepatocellular glutathione contentin control rats at time 0 and after 20 day diet treatment.Hepatocellular O₂ Reduced consumption glutathione Mitochondrial (umolesO₂/min (nmoles × 10⁶ membrane per 10⁷ cells) cells) potential 1A)Control rats 480 ± 60 48 ± 5 100% at time 0 1B) Control rats after 360 ±45 36 ± 4 68% a 20-day diet 2) Treated rats (NOE) 368 ± 52 36 ± 6 70% 3)Treated rats (NOPE) 408 ± 62 41 ± 5 81% 4) Treated rats (NOE + PA) 412 ±58 43 ± 6 82% 5) Treated rats (NOE + LPA) 409 ± 63 43 ± 8 83% 6) Treatedrats (NOPE + B.F.) 421 ± 51 45 ± 5 88% 7) Treated rats (B.F.) 366 ± 4138 ± 5 71%

When the membrane fluidity of the ghost erythrocytes and plasmaplatelets is measured, TMA-DPH in accordance with the method describedby Caimi F. et al., 1999, Thromb. Hoemost., 82 pp. 149, is used as thefluorescent probe.

Malonyldialdehyde is assayed in accordance with the procedure describedby K. Yagi et al., 1982, in “Lipid Peroxides in Biology and Medicine”,Academic Press, New York, 99. 324-340.

Hepatocellular O₂ consumption, mitochondrial membrane potential andreduced glutathione content are assayed in accordance with the methodsdescribed by T. M. Hagen et al., 1999, FASEB J., 13, 99. 411.

The data set out in Tables I, II, III and IV demonstrate thatadministration of compositions containing the active components (NOPE;NOE+PA; NOE+LPA; NOPE+B.F.):

-   -   1) restores the membrane fluidity of ghost and platelets;    -   2) improves the antioxidant defences of plasma, liver, brain and        heart;    -   3) limits excessive increases in body weight;    -   4) limits excessive increases in plasma cholesterol and        triglyceride levels;    -   5) improves the functionality of the mitochondria.

These effects, obtainable by oral administration of the formulationsprepared in accordance with the invention (NOPE; NOE+PA; NOE+LPA;NOPE+B.F.), are always statistically significant. It is important tonote that no statistically significant benefit can be obtained byadministering equivalent oral doses of N-oleoyl-ethanolamines as such.

The data set out above demonstrate the surprising synergy of actionobserved between NAPE and/or NAE+PA and the various bioflavonoidmolecules; the therapeutic results obtainable by administering the“phosphobioflavonic complexes” of NAPE (see data set out in Tables I,II, III and IV) and NAE plus PA and/or LPA are always far higher thanthe sum of the benefits obtainable with single separate administrationsof equivalent doses of NAPE (or NAE) and bioflavonoids.

In all the diet treatment tests carried out on man, the effectsobtainable by orally the formulations claimed by the invention (NAPE;NAE+PA; NAE+LPA; NAPE+B.F. and NAE plus PA and/or LPA+B.F.) alwaysprovided highly significant results and advantages, both in preventingbiological signs of aging (improvement in mitochondrial activity, bettermembrane fluidity, improvement in plasma antioxidant defences, andlimited weight increase) and improving the clinical parameters tested inrelation to prevention of aging, and many of the metabolic disordersassociated therewith. It is noteworthy that also in humans nosignificant benefit can be obtained by administering equivalent oraldoses of N-oleoyl-ethanolamine as such.

1. Pharmaceutical, cosmetic and dietetic compositions and functionalfoods, constituted by: A) phospholipid mixtures containingN-acyl-phosphatidyl-ethanolamines (NAPEs); and/or B) phospholipidsmixtures containing N-acyl-ethanolamines (NAEs) together withphosphatidic acids (PAs) and/or lysophosphatidic acids (LPAs), with theproviso that said N-acyl-phosphatidyl-ethanolamines (NAPEs) do notinclude N-oleoyl-phosphatidyl-ethanolamine.
 2. Compositions as claimedin claim 1 wherein the phospholipid mixtures are constituted bycomplexes of one or more bioflavones with NAPE or NAE plus PA and/orLPA.
 3. Compositions as claimed in claim 1, also containing amphiphilicsurfactants.
 4. Functional foodstuffs containing the compositionsclaimed in claim
 1. 5. The use of phospholipid mixtures containing A)N-acyl-phosphatidyl-ethanolamines (NAPEs); and/or B) phospholipidmixtures containing N-acyl-ethanolamines (NAEs) together withphosphatidic acids (PAs) and/or lysophosphatidic acids (LPAs). for thepreparation of medicaments having anorexic activity or of medicaments orfoodstuffs for the treatment of aging, obesity and excess weight;diabetes; cerebro-degenerative disorders such as Alzheimer's disease,Parkinson's disease and senile dementia; stress, depression; tumours;menopaulsal syndromes; osteoporosis; prostate hypertrophy; skin aging;panniculopathy (cellulitis) and alopecia.
 6. Phosphobioflavoniccomplexes of bioflavonoids with phospholipids active components selectedfrom N-acyl-phosphatidyl-ethanolamines (NAPE) and/orN-acyl-ethanolamines (NAE) plus Phosphatidic Acid (PA) and/orN-acyl-ethanolamines plus Lysophosphatidic acid (LPA).